Tunnel engineering for modulating the substrate preference in cytochrome P450BsβHI

Bioresources and Bioprocessing

Ruipeng An¹  Luo Liu¹  Zhongyu Li¹  Meng Qin¹  Kaili Nie¹  Meng Wang¹  Fang Wang¹  Shuaiqi Meng¹  Mehdi D. Davari²  Ulrich Schwaneberg²  Yu Ji² 

[1] Beijing Bioprocess Key Laboratory, Beijing University of Chemical Technology;Institute of Biotechnology, RWTH Aachen University;

关键词: Tunnel engineering;    Substrate preference;    Cytochrome P450BsβHI;    α-Alkene biosynthesis;    Rational design;   

DOI  :  10.1186/s40643-021-00379-1

来源: DOAJ

【 摘 要 】

Abstract An active site is normally located inside enzymes, hence substrates should go through a tunnel to access the active site. Tunnel engineering is a powerful strategy for refining the catalytic properties of enzymes. Here, P450BsβHI (Q85H/V170I) derived from hydroxylase P450Bsβ from Bacillus subtilis was chosen as the study model, which is reported as a potential decarboxylase. However, this enzyme showed low decarboxylase activity towards long-chain fatty acids. Here, a tunnel engineering campaign was performed for modulating the substrate preference and improving the decarboxylation activity of P450BsβHI. The finally obtained BsβHI-F79A variant had a 15.2-fold improved conversion for palmitic acid; BsβHI-F173V variant had a 3.9-fold improved conversion for pentadecanoic acid. The study demonstrates how the substrate preference can be modulated by tunnel engineering strategy.

【 授权许可】

Unknown